Detection and enrichment of antigen-specific CD4+ and CD8+ T cells based on cytokine secretion

The Cytokine Secretion Assay is an innovative method for analysing and enriching live cytokine-secreting cells. In this assay, a cytokine affinity matrix is built on the cell plasma membrane, which traps cytokines produced by the cell in response to specific stimuli. The specifically bound cytokine is then detected, and the cells optionally enriched, using fluorochrome-conjugated cytokine-specific antibodies and magnetic microbeads. This method allows extremely detailed phenotyping of live cells and the detection of cytokine responses at very low frequencies. Here, the latest cell staining and separation procedures are reviewed, with particular reference to the best application of the technology and troubleshooting in a variety of different situations.